First Report of Pomegranate Wilt Caused by Ceratocystis fimbriata in Yunnan, China

Pomegranate (Punica granatum Linn.) is an important fruit crop in Yunnan Province, China. Recently, older pomegranate bushes in Mengzi County, Yunnan began dying. Initial symptoms were yellowing and wilting of leaves on one to several branches, followed by sudden death of the bush within 3 to 4 weeks. Roots of diseased bushes appeared brown to black, and irregularly shaped lesions were observed when the bark was removed. A species of Ceratocystis was consistently isolated from discolored roots, stem, and branch tissues from wilted bushes on potato dextrose agar (PDA) and was identified as Ceratocystis fimbriata Ellis & Halst, based on the morphology of perithecia, ascospores, conidia, and conidiophores. Perithecia were black with a globose base (130 to 300 μm) and a long neck (450 to 800 μm). Ascospores exuded from the apex of the perithecium neck in a long coil and were small, hyaline, and hat-shaped (3.8 to 5.0 μm long × 2.3 to 4.0 μm wide). Conidiophores were septate and hyaline to dark greenish brown. Hyaline conidia, 8 to 17 μm long × 6 to 15 μm wide, were usually produced in chains of 10 or more. Thick-walled endoconidia were globose to oval, olive brown, and 8 to 20 μm in diameter. Because of the increasing occurrence of the disease, surveys of the main pomegranate production areas in Mengzi County, including Xinan, Duofale, Caoba, Hongzhai, and Shilipu townships, were conducted from 10 to 20 August 2002. The disease was detected in 17 of 50 plantings surveyed. Disease was more severe in older plantings than in younger plantings. Disease incidence was 1% in 1- to 5-year-old bushes, 3.6% in 6- to 10-year-old bushes, and 6% in bushes more than 10 years old. Scolytid beetles were occasionally found on bushes, but we were unable to isolate the fungus from them as has been reported (1). Inoculations with an isolate of C. fimbrata were made by inserting mycelium with perithecia from 12-day-old cultures growing on PDA into root wounds made with a sterile scalpel on five pomegranate plants and then covering the wounds with Parafilm. Sterile medium was placed in an equal number of wounded bushes to serve as controls. Fourteen days later, symptoms began to appear in two bushes, and 5 days later, all bushes exhibited symptoms. No symptoms were observed on control bushes. The first visible symptom was a small area of blackened tissue near the point of inoculation. Lesions expanded slowly, but they expanded more rapidly upward than downward. The fungus was reisolated on PDA from roots of all artificially inoculated bushes. C. fimbriata has been previously reported as the cause of pomegranate wilt in India (2); however, to our knowledge, this is the first report of C. fimbriata on pomegranate in China. Because environmental conditions which favor the pathogen (temperatures ranging from 18 to 30°C and frequent rains) typically occur in many areas during late spring and summer, the disease has the potential to seriously impact pomegranate production in China. References: (1) Y. M. Somasekhara. Plant Dis. 83:400, 1999. (2) Y. M. Somasekhara, et al. Res. Crops 1(1):63, 2000.

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Ginger wilt and rot disease caused by Ceratocystis fimbriata in China

Plant Disease ◽

10.1094/pdis-08-20-1647-pdn ◽

2020 ◽

Author(s):

Ruiqi Zhang ◽

Kecheng Xu ◽

Xue Li ◽

Yang Gao ◽

Yuexian Sun ◽

...

Keyword(s):

Yunnan Province ◽

Disease Incidence ◽

Internal Transcribed Spacers ◽

Therapeutic Drug ◽

Malt Extract ◽

Sterile Water ◽

Ceratocystis Fimbriata ◽

Single Spore ◽

Initial Symptoms ◽

Rot Disease

Ginger (Zingiber officinale Rosc.) is an herb that has been grown in China for more than 2500 years. It can be used as both a spice and a therapeutic drug. In July 2013, ginger plants were found to have wilting symptoms and yellowing leaves with netrotics leaf tips in a farm in Kunming city of Yunnan province (25. 02 N; 102.42 E), southwest China, and we also found gray-black lesion on the surface of the harvest gingers in a market in Kunming. Initial symptoms on harvest gingers appeared as gray-black mycelia growth on the surface of the harvested ginger, which enlarged and extended internally. Carrot baiting was used to isolate the pathogen from rotted gingers and diseased ginger leaves (Moller and Devay. 1968). After two weeks, spores developing from perithecia on the carrot pieces were transferred to malt extract agar (MEA) and incubate at 25°C constant-temperature incubator. Six single-spore isolates (ZOR-1 to ZOR-6) were obtained, the isolates were stored in 15% glycerol at -80°C refrigerator in State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan Agricultural University. Cultures varied in color from white to brownish green to brown. N = 50 for all measurements. Blackish brown, globose perithecia (131.9 to 186.0 μm × 138.5 to 188.3 μm) with a long black neck (400.2 to 644.7 μm) were immersed, partially embedded or superficial on the substrate. Ascospores were globose or had a “hat-like” morphology typical of Ceratocystis fimbriata, and were 4.0 to 5.3 μm × 4.8 to 6.2 μm. Endoconidia were cylindrical and clavate (2.9 to 7.4 μm × 7.5 to 32.8 μm), conidia were barrel-shaped (4.4 to 10.4 μm× 6.2 to 12.9 μm), and chlamydospores were smooth, blackish brown, ovoid or obpyriform (8.42 to 12.21 μm × 10.47 to 17.65 μm) (Webster and Butler. 1967; Engelbrecht and Harrington. 2005). Genomic DNA was extracted from two isolates (ZOR-1, ZOR-2) using the CTAB method (Lee and Taylor 1990). The internal transcribed spacers (ITS) region of rDNA was amplified using primers ITS1F/ITS4 (Thorpe et al. 2005). The nucleotide sequences of ZOR-1 and ZOR-2 (GenBank accessions KJ511490 and KJ511491) were 100% homologous to those of the isolates of C. fimbriata from diseased Cucumis sativus L. and Punica granatum L. in China (GenBank accessions MH535909 and KT963159). Thus, the pathogen was identified as C. fimbriata. Pathogenicity tests were made on fresh ginger rhizomes in laboratory, the pathogen was cultured for 14 days on MEA (ZOR-1, ZOR-2), which were washed with sterilized water and the resulting spore suspensions diluted to 1.0 × 106 spores/ml . Wounds (0.5 × 0.5 cm) were made on the surface of healthy mature ginger rhizomes by scraping with a sterile scalpel, then treated with a 100 ul spore suspension. Control ginger rhizomes were coated sterile water. Ginger rhizomes were stored at room temperature. Each treatment was performed in triplicate. After 5 days, grey-black mycelia developed on the rhizome surface, becoming a visible black mould after 1 week. We reisolated the pathogen from infected tissues, but not from the controls. In the greenhouse, 20ml of 1.0 × 106 spores/ml suspensions from isolates ZOR-1 and ZOR-2, or sterile water were injected into two-month- old ginger seedlings in triplicate. The inoculated site on the stem turned black in 5 days. 6 weeks after inoculation, the inoculated plants developed yellowing leaves and wilting symptoms. The same fungus was re-isolated from inoculated plants, but not from the controls. According to Koch’s Postulation, the inoculated strains of ZOR-1 and ZOR-2 were the pathogens causing ginger wilt and rot disease. To the best of our knowledge, ginger is a new host plant of Ceratocystis fimbriata from China. In recent years, we have found that this disease incidence was approxmiatelt 5 to 10% of the farmland and 5 to 15% of the stored condition respectively in Yunnan Province. If not prevented ginger production in China will be affected.

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First Report of Clover Proliferation Group Phytoplasmas (16SrVI-A) Associated with Purple Top Diseased Potatoes (Solanum tuberosum) in China

Plant Disease ◽

10.1094/pdis-03-11-0152 ◽

2011 ◽

Vol 95 (7) ◽

pp. 871-871 ◽

Cited By ~ 4

Author(s):

M. Cheng ◽

J. Dong ◽

P. J. Laski ◽

Z. Zhang ◽

J. H. McBeath

Keyword(s):

Nested Pcr ◽

Disease Incidence ◽

Restriction Enzymes ◽

Potato Production ◽

First Report ◽

Growing Seasons ◽

Pcr Product ◽

Disease Surveys ◽

Production Areas ◽

Inner Mongolia Autonomous Region

Phytoplasma diseases on potatoes are not well understood and have gone largely undetected in China. During the growing seasons of 2005 through 2010, potato disease surveys were conducted in seed and commercial fields in Yunnan Province. Samples were also harvested from three seed potato production areas in the Inner Mongolia Autonomous Region in 2007 and 2010. Disease incidence in these fields ranged from 15 to 85%. Plants displayed symptoms of branch proliferation, aerial tuber formation, upward rolling yellowish and purplish apical leaves, and extremely short stolen or chain tubers (irregular-shaped tubers). Total DNA from 250 samples was extracted from the leaves, stems, and roots of symptomatic and asymptomatic plants. A nested PCR was performed by using primer pair P1/P7 followed by R16F2n/R16R2 to detect the presence of phytoplasmas (1,3). An approximate 1.2-kb PCR product was amplified from symptomatic plants but not from asymptomatic plants. Restriction fragment length polymorphism (RFLP) patterns were analyzed by digesting the 1.2-kb amplicon singly with restriction enzymes AluI, BfaI, MseI, HhaI, HinfI, HpaII, KpnI, RsaI, and TaqI. The RFLP patterns of 120 of the 250 samples matched patterns of the clover proliferation (CP) group (16SrVI) subgroup A (16SrVI-A) phytoplasma (1). In addition, the nested PCR product of P1A/P7A (2) following P1/P7 amplification was cloned and sequenced (GenBank Accession No. HQ609490). Nucleotide sequences were analyzed by iPhyClassifier software (4), confirming the relationship of this phytoplasma to ‘Candidatus Phytoplasma trifolii’ with RFLP patterns identical to group 16SrVI-A. To our knowledge, this is the first report of the CP group phytoplasmas associated with purple top diseased potatoes in China. References: (1) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (2) I.-M. Lee et al. Int. J. Syst. Evol. Microbiol 54:337, 2004. (3) C. D. Smart et al. Appl. Environ. Microbiol. 62:2988, 1996. (4) Y. Zhao et al. Int. J. Syst. Evol. Microbiol. 59:2582, 2009.

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First Report of Anthracnose of Papaya (Carica papaya L.) Caused by Colletotrichum siamense in China

Plant Disease ◽

10.1094/pdis-10-20-2154-pdn ◽

2021 ◽

Author(s):

Yujie Zhang ◽

Wenxiu Sun ◽

Ping Ning ◽

Tangxun Guo ◽

SuiPing Huang ◽

...

Keyword(s):

Carica Papaya ◽

Disease Incidence ◽

Aerial Mycelium ◽

Postharvest Disease ◽

Distilled Water ◽

First Report ◽

Surface Samples ◽

Initial Symptoms ◽

Colletotrichum Siamense ◽

Pathogenicity Tests

Papaya (Carica papaya L.) is a rosaceous plant widely grown in China, which is economically important. Anthracnose caused by Colletotrichum sp. is an important postharvest disease, which severely affects the quality of papaya fruits (Liu et al., 2019). During April 2020, some mature papaya fruits with typical anthracnose symptoms were observed in Fusui, Nanning, Guangxi, China with an average of 30% disease incidence (DI) and over 60% DI in some orchards. Initial symptoms of these papayas appeared as watery lesions, which turned dark brown, sunken, with a conidial mass appearing on the lesions under humid and warm conditions. The disease severity varied among fruits, with some showing tiny light brown spots, and some ripe fruits presenting brownish, rounded, necrotic and depressed lesions over part of their surface. Samples from two papaya plantations (107.54°E, 22.38°N) were collected, and brought to the laboratory. Symptomatic diseased tissues were cut into 5 × 5 mm pieces, surface sterilized with 2% (v/v) sodium hypochlorite for 1 minute, and rinsed three times with sterilized water. The pieces were then placed on potato dextrose agar (PDA). After incubation at 25°C in the dark for one week, colonies with uniform morphology were obtained. The aerial mycelium on PDA was white on top side, and concentric rings of salmon acervuli on the underside. A gelatinous layer of spores was observed on part of PDA plates after 7 days at 28°C. The conidia were elliptical, aseptate and hyaline (Zhang et al., 2020). The length and width of 60 conidia were measured for each of the two representative isolates, MG2-1 and MG3-1, and these averaged 13.10 × 5.11 μm and 14.45 × 5.95 μm. DNA was extracted from mycelia of these two isolates with the DNA secure Plant Kit (TIANGEN, Biotech, China). The internal transcribed spacer (ITS), partial actin (ACT), calmodulin (CAL), chitin synthase (CHS), β-tubulin 2 (TUB2) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) regions were amplified by PCR and sequenced. The sequences were deposited into GenBank with accessions MT904003, MT904004, and MT898650 to MT898659. BLASTN analyses against the GenBank database showed that they all had over 99% identity to the type strain of Colletotrichum siamense isolate ICMP 18642 (GenBank accession numbers JX010278, GQ856775, JX009709, GQ856730, JX010410, JX010019) (Weir et al., 2012). A phylogenetic tree based on the combined ITS, ACT, CAL, CHS, TUB2 and GAPDH sequences using the Neighbor-joining algorithm also showed that the isolates were C. siamense. Pathogenicity tests were conducted on 24 mature, healthy and surface-sterilized papaya fruits. On 12 papaya fruits, three well separated wounded sites were made for inoculation, and for each wounded site, six adjacent pinhole wounds were made in a 5-mm-diameter circular area using a sterilized needle. A 10 µl aliquot of 1 × 106 conidia/ml suspension of each of the isolates (MG2-1 and MG3-1) was inoculated into each wound. For each isolate, there were six replicate fruits. The control fruits were inoculated with sterile distilled water. The same inoculation was applied to 12 non-wound papaya fruits. Fruits were then placed in boxes which were first washed with 75% alcohol and lined with autoclaved filter paper moistened with sterilized distilled water to maintain high humidity. The boxes were then sealed and incubated at 28°C. After 10 days, all the inoculated fruits showed symptoms, while the fruits that were mock inoculated were without symptoms. Koch's postulates were fulfilled by re-isolation of C. siamense from diseased fruits. To our knowledge, this is the first report of C. siamense causing anthracnose of papaya in China. This finding will enable better control of anthracnose disease caused by C. siamense on papaya.

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First Report of a Leaf Spot on Snow Lotus Caused by Alternaria carthami in China

Plant Disease ◽

10.1094/pdis-92-2-0318b ◽

2008 ◽

Vol 92 (2) ◽

pp. 318-318

Author(s):

S. Zhao ◽

G. Xie ◽

H. Zhao ◽

H. Li ◽

C. Li

Keyword(s):

Leaf Spot ◽

Disease Incidence ◽

Leaf Spot Disease ◽

Tianshan Mountain ◽

Causal Organism ◽

Saussurea Involucrata ◽

First Report ◽

Spot Disease ◽

Initial Symptoms ◽

Snow Lotus

Snow lotus (Saussurea involucrata Karel. & Kir. ex Sch. Bip.) is an economically important medicinal herb increasingly grown in China in recent years. In June of 2005, a leaf spot disease on commercially grown plants was found in the QiTai Region, south of the Tianshan Mountain area of Xinjiang, China at 2,100 m above sea level. Disease incidence was approximately 60 to 70% of the plants during the 2006 and 2007 growing seasons. Initial symptoms appeared on older leaves as irregularly shaped, minute, dark brown-to-black spots, with yellow borders on the edge of the leaflet blade by July. As the disease progressed, the lesions expanded, causing the leaflets to turn brown, shrivel, and die. A fungus was consistently isolated from the margins of these lesions on potato dextrose agar. Fifty-eight isolates were obtained that produced abundant conidia in the dark. Conidia were usually solitary, rarely in chains of two, ellipsoid to obclavate, with 6 to 11 transverse and one longitudinal or oblique septum. Conidia measured 60 to 80 × 20 to 30 μm, including a filamentous beak (13 to 47 × 3.5 to 6 μm). According to the morphology, and when compared with the standard reference strains, the causal organism of leaf spot of snow lotus was identified as Alternaria carthami (1,4). Pathogenicity of the strains was tested on snow lotus seedlings at the six-leaf stage. The lower leaves of 20 plants were sprayed until runoff with conidial suspensions of 1 × 104 spores mL–1, and five plants sprayed with sterile distilled water served as controls. All plants were covered with a polyethylene bag, incubated at 25°C for 2 days, and subsequently transferred to a growth chamber at 25°C with a 16-h photoperiod. Light brown lesions developed within 10 days on leaflet margins in all inoculated plants. The pathogen was reisolated from inoculated leaves, and isolates were deposited at the Key Oasis Eco-agriculture Laboratory of Xinjiang Production and Construction Group, Xinjiang and the Institute of Biotechnology, Zhejiang University. No reports of a spot disease caused by A. carthami on snow lotus leaves have been found, although this pathogen has been reported on safflower in western Canada (3), Australia (2), India (1), and China (4). To our knowledge, this is the first report of a leaf spot caused by A. carthami on snow lotus in China. References: (1) S. Chowdhury. J. Indian Bot. Soc. 23:59, 1944. (2) J. A. G. Irwin. Aust. J. Exp. Agric. Anim. Husb. 16:921, 1976. (3) G. A. Petrie. Can. Plant Dis. Surv. 54:155, 1974. (4) T. Y. Zhang. J. Yunnan Agric. Univ.17:320, 2002.

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First Report of a Leaf Spot Disease of Bells-of-Ireland (Moluccella laevis) Caused by Cercospora apii in California

Plant Disease ◽

10.1094/pdis.2003.87.2.203a ◽

2003 ◽

Vol 87 (2) ◽

pp. 203-203

Author(s):

S. T. Koike ◽

S. A. Tjosvold ◽

J. Z. Groenewald ◽

P. W. Crous

Keyword(s):

Leaf Spot ◽

Sequence Data ◽

Disease Incidence ◽

Leaf Spot Disease ◽

Conidial Suspension ◽

Internal Transcribed Spacer Region ◽

First Report ◽

Spot Disease ◽

Leaf Spots ◽

Initial Symptoms

Bells-of-Ireland (Moluccella laevis) (Lamiaceae) is an annual plant that is field planted in coastal California (Santa Cruz County) for commercial cutflower production. In 2001, a new leaf spot disease was found in these commercially grown cutflowers. The disease was most serious in the winter-grown crops in 2001 and 2002, with a few plantings having as much as 100% disease incidence. All other plantings that were surveyed during this time had at least 50% disease. Initial symptoms consisted of gray-green leaf spots. Spots were generally oval in shape, often delimited by the major leaf veins, and later turned tan. Lesions were apparent on both adaxial and abaxial sides of the leaves. A cercosporoid fungus having fasciculate conidiophores, which formed primarily on the abaxial leaf surface, was consistently associated with the spots. Based on morphology and its host, this fungus was initially considered to be Cercospora molucellae Bremer & Petr., which was previously reported on leaves of M. laevis in Turkey (1). However, sequence data obtained from the internal transcribed spacer region (ITS1, ITS2) and the 5.8S gene (STE-U 5110, 5111; GenBank Accession Nos. AY156918 and AY156919) indicated there were no base pair differences between the bells-of-Ireland isolates from California, our own reference isolates of C. apii, as well as GenBank sequences deposited as C. apii. Based on these data, the fungus was subsequently identified as C. apii sensu lato. Pathogenicity was confirmed by spraying a conidial suspension (1.0 × 105 conidia/ml) on leaves of potted bells-of-Ireland plants, incubating the plants in a dew chamber for 24 h, and maintaining them in a greenhouse (23 to 25°C). After 2 weeks, all inoculated plants developed leaf spots that were identical to those observed in the field. C. apii was again associated with all leaf spots. Control plants, which were treated with water, did not develop any symptoms. The test was repeated and the results were similar. To our knowledge this is the first report of C. apii as a pathogen of bells-of-Ireland in California. Reference: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. Cornell University Press, Ithaca, New York, 1954.

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First Report of Curvularia aeria on Etlingera linguiformis from Nagaland, India

Plant Disease ◽

10.1094/pdis-01-14-0060-pdn ◽

2014 ◽

Vol 98 (11) ◽

pp. 1580-1580 ◽

Cited By ~ 2

Author(s):

C. Kithan ◽

L. Daiho

Keyword(s):

Leaf Surface ◽

Agricultural Research ◽

Disease Incidence ◽

Indian Agricultural Research Institute ◽

Mountain Range ◽

Pathogenicity Test ◽

Distilled Water ◽

Conidial Suspension ◽

First Report ◽

Initial Symptoms

Etlingera linguiformis (Roxb.) R.M.Sm. of Zingiberaceae family is an important indigenous medicinal and aromatic plant of Nagaland, India, that grows well in warm climates with loamy soil rich in humus (1). The plant rhizome has medicinal benefits in treating sore throats, stomachache, rheumatism, and respiratory complaints, while its essential oil is used in perfumery. A severe disease incidence of leaf blight was observed on the foliar portion of E. linguiformis at the Patkai mountain range of northeast India in September 2012. Initial symptoms of the disease are small brown water soaked flecks appearing on the upper leaf surface with diameter ranging from 0.5 to 3 cm, which later coalesced to form dark brown lesions with a well-defined border. Lesions often merged to form large necrotic areas, covering more than 90% of the leaf surface, which contributed to plant death. The disease significantly reduces the number of functional leaves. As disease progresses, stems and rhizomes were also affected, reducing quality and yield. The diseased leaf tissues were surface sterilized with 0.2% sodium hypochlorite for 2 min followed by rinsing in sterile distilled water and transferred into potato dextrose agar (PDA) medium. After 3 days, the growing tips of the mycelium were transferred to PDA slants and incubated at 25 ± 2°C until conidia formation. Fungal colonies on PDA were dark gray to dark brown, usually zonate; stromata regularly and abundantly formed in culture. Conidia were straight to curved, ellipsoidal, 3-septate, rarely 4-septate, middle cells broad and darker than other two end cells, middle septum not median, smooth, 18 to 32 × 8 to 16 μm (mean 25.15 × 12.10 μm). Conidiophores were terminal and lateral on hyphae and stromata, simple or branched, straight or flexuous, often geniculate, septate, pale brown to brown, smooth, and up to 800 μm thick (2,3). Pathogen identification was performed by the Indian Type Culture Collection, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi (ITCC Accession No. 7895.10). Further molecular identity of the pathogen was confirmed as Curvularia aeria by PCR amplification and sequencing of the internal transcribed spacer (ITS) regions of the ribosomal DNA by using primers ITS4 and ITS5 (4). The sequence was submitted to GenBank (Accession No. MTCC11875). BLAST analysis of the fungal sequence showed 100% nucleotide similarity with Cochliobolus lunatus and Curvularia aeria. Pathogenicity tests were performed by spraying with an aqueous conidial suspension (1 × 106 conidia /ml) on leaves of three healthy Etlingera plants. Three plants sprayed with sterile distilled water served as controls. The first foliar lesions developed on leaves 7 days after inoculation and after 10 to 12 days, 80% of the leaves were severely infected. Control plants remained healthy. The inoculated leaves developed similar blight symptoms to those observed on naturally infected leaves. C. aeria was re-isolated from the inoculated leaves, thus fulfilling Koch's postulates. The pathogenicity test was repeated twice. To our knowledge, this is the first report of the presence of C. aeria on E. linguiformis. References: (1) M. H. Arafat et al. Pharm. J. 16:33, 2013. (2) M. B. Ellis. Dematiaceous Hyphomycetes. CMI, Kew, Surrey, UK, 1971. (3) K. J. Martin and P. T. Rygiewicz. BMC Microbiol. 5:28, 2005. (4) C. V. Suberamanian. Proc. Indian Acad. Sci. 38:27, 1955.

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Sugarcane Mosaic Distribution, Incidence, Increase, and Spatial Pattern in Louisiana

Plant Disease ◽

10.1094/pdis-01-19-0099-re ◽

2019 ◽

Vol 103 (8) ◽

pp. 2051-2056

Author(s):

J. L. Rice ◽

J. W. Hoy ◽

M. P. Grisham

Keyword(s):

Spatial Pattern ◽

Disease Incidence ◽

Aphid Transmission ◽

Field Surveys ◽

Reverse Transcription Pcr ◽

Single Leaf ◽

Disease Surveys ◽

Low Incidence ◽

Mosaic Distribution ◽

Production Areas

Sugarcane mosaic is a historically important disease in Louisiana currently caused by sorghum mosaic virus (SrMV). Successful breeding for resistance reduced the disease to low incidence in commercial cultivars. However, mosaic was detected in experimental clone evaluations at multiple locations, leading to uncertainty concerning the current distribution and incidence in the state. Field surveys were conducted from 2016 to 2018 in breeding program yield trials and experimental clone seed cane increase fields. Mosaic symptomatic plants were observed in a newly released cultivar, HoCP 09-804, in three of five production areas, with incidences ranging from 0 to 10%. Mosaic also was observed in nine additional experimental clones. Single leaf samples were tested for SrMV using reverse transcription PCR. All symptomatic samples and a low percentage (0.3%) of asymptomatic samples tested positive for SrMV, confirming that it continues to be the causal species. Runs analysis detected aggregation of infected plants within at least 70% of rows in 94% of surveyed fields. The spatial pattern and geographical distribution of disease incidence suggested that infected seed cane was the source of the disease. Surveys conducted in the same fields of HoCP 09-804 through two subsequent crops detected disease incidence increases in some fields and decreases in the others in first ratoon, but observed incidence was lower compared with plant cane in all fields in second ratoon. The results indicated that disease increase owing to aphid transmission did not occur under the prevailing conditions.

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First Report of Bacterial Wilt Caused by Ralstonia solanacearum on Plectranthus amboinicus in Martinique

Plant Disease ◽

10.1094/pdis-12-20-2622-pdn ◽

2021 ◽

Author(s):

Peninna Deberdt ◽

Gilles Cellier ◽

Régine Coranson-Beaudu ◽

Mathis Delmonteil--Girerd ◽

Joanye Canguio ◽

...

Keyword(s):

Bacterial Wilt ◽

Ralstonia Solanacearum ◽

Disease Incidence ◽

Bacterial Suspension ◽

Diagnostic Pcr ◽

Tomato Plants ◽

First Report ◽

Initial Symptoms ◽

Plectranthus Amboinicus ◽

Negative Controls

Plectranthus amboinicus, commonly known as Gwo ten in the French West Indies (Martinique), is a semi-succulent perennial plant of the Lamiaceae family. This aromatic plant wich is widespread naturally throughout the tropics is of economic importance because of the therapeutic and nutritional properties attributed to its natural phytochemical compounds wich are highly valued in the pharmaceutical industry. In March 2019, wilted P. amboinicus plants intercropped with tomato plants (cv. Heatmaster) in order to reduce the insect-pest damages on tomato, were observed in a field located at the CIRAD experimental station in Lamentin, Martinique (14.663194 N, -60.999167 W). Average disease incidence of 65.74% was recorded on P. amboinicus, in 3 plots with an area of 22.04 m2. The initial symptoms observed were irregular, black, necrotic lesions on leaves. After 10 days, plants wilted and black stripes were observed on stems. Within 4 weeks, more than 50% of plants were fully wilted. Longitudinal stem sections of the wilted plants showed brown vascular discoloration. The cut stems of the wilted plants released a whitish bacterial ooze in water. In all, 108 stem sections were collected, surface disinfected with 70% ethanol and each was crushed in 2 mL of Tris-buffer, then processed for bacterial isolation by plating on modified Semi-Selective Medium from South Africa SMSA (Engelbrecht 1994). Typical Ralstonia solanacearum colonies grew on SMSA medium for 100 of the 108 samples after incubation for 48h at 28°C and were identified as Ralstonia solanacearum using diagnostic PCR with 759/760 primers (Opina et al. 1997). A phylotype-specific multiplex PCR (Fegan and Prior 2005) classified all the strains in R. solanacearum Phylotype IIA. A subset of 11 strains was selected for sequevar identification. All the strains were identified as sequevar I-39 (100% nucleotide identity with strain ANT92 - Genbank accession EF371828), by partial egl sequencing (Fegan and Prior 2005) (GenBank Accession Nos. MT314067 to MT314077). This sequevar has been reported to be widespread in the Caribbean and tropical America on vegetable crops (particularly on tomato), but not on P. amboinicus (Deberdt et al. 2014; Ramsubhag et al. 2012; Wicker et al. 2007). To fulfil Koch’s postulates, a reference strain, isolated from diseased P. amboinicus (CFBP 8733, Phylotype IIA/sequevar 39), was inoculated on 30 healthy P. amboinicus plants. A common tomato cultivar grown in Martinique (cv. Heatmaster) was also inoculated on 30 plants with the same bacterial suspension. Three-weeks-old plants of both crops grown in sterilized field soil were inoculated by soil drenching with 20 ml of a calibrated suspension (108 CFU/mL). P. amboinicus and tomato plants drenched with sterile water served as a negative controls. Plants were grown in a fully controlled environment at day/night temperatures of 30–26°C ± 2°C under high relative humidity (80%). The P. amboinicus plants started wilting 9 days after inoculation, and within four weeks 60% of the P. amboinicus plants had wilted. The tomato plants started wilting 5 days after inoculation with 62% of wilted plants within four weeks. R. solanacearum was recovered from all symptomatic plants on modified SMSA medium. No symptoms were observed and no R. solanacearum strains were isolated from negative controls plants. To our knowledge, this is the first report of R. solanacearum causing bacterial wilt on Gwo ten (P. amboinicus) in Martinique. The importance of this discovery lies in the reporting of an additional host for R. solanacearum, which can be associated with other crops as tomato crop in order to reduce the abundance of insect-pests. Further studies need to be conducted to assess the precise distribution of bacterial wilt disease on P. amboinicus in Martinique and to develop a plan of action avoiding its association with R. solanacearum host crops as tomato for reducing epidemic risk.

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First Report of Meloidogyne incognita Infecting Corn in Western Spain

Plant Disease ◽

10.1094/pdis-07-12-0674-pdn ◽

2013 ◽

Vol 97 (5) ◽

pp. 694-694 ◽

Cited By ~ 1

Author(s):

J. López-Robles ◽

P. García-Benavides ◽

G. Sacristán-Pérez-Minayo

Keyword(s):

Crop Yields ◽

Disease Incidence ◽

Nematode Population ◽

Root Knot Nematode ◽

Root Tips ◽

First Report ◽

North Carolina State University ◽

Disease Surveys ◽

Western Spain ◽

Corn Plants

During the 2009 to 2010 corn-growing (Zea mays L. cv. Alexandria) seasons, severely stunted and yellowing corn plants in several commercial fields at Aldearrubia (Salamanca Province) were observed in western Spain. The disease incidence ranged from 80 to 100%. Early symptoms consisted of severely reduced growth of the plants coupled with extensive leaf yellowing. Occurrence of the disease was estimated to cause near complete loss of the crop yields since the corn produced in affected fields was unmarketable. Disease surveys revealed high parasitism in the main and feeder roots and a large soil population of the root-knot nematode Meloidogyne spp. The nematode population was extracted and quantified from soil and root samples according to Barker (1). It was identified as the southern root-knot nematode M. incognita race 1, by female perineal pattern, host-differential test, and multiplex PCR using forward primers H-18S, CF-ITS, I-ITS, and reverse primer HCFI-28S (3,4). ITS products cloned and assayed using the ABI PRISM 3100 Genetic Analyzer (Applied Biosystems, Salamanca, Spain) were subjected to a database search using BLAST (National Centre for Biotechnology Information) to confirm the identification. These sequences exhibited 99.0% similarity with that of an M. incognita isolate from France (GenBank Accession No. AF402309.1). M. incognita was found in 80% of soil samples collected from the areas where the disease was observed and 83.5% of root samples with nematode population densities ranging from 26 to 269 eggs and second-stage juveniles (J2s) per 100 cm3 of soil and 234 to 1,634 eggs and J2s per 5 g of fresh roots. In glasshouse proofs of pathogenicity, a mix of 1,500 eggs and J2s of these populations of M. incognita were inoculated in 20-cm-diameter pots with 10 replicates with a single pregerminated seed of corn cv. Alexandria as host plant; another 10 replicates without inoculation were established as control plants. After 6 weeks, all plants inoculated were severely stunted and yellowing; infected roots showed galls on root tips and secondary feeder roots. Galling of root tips that cause stubby root symptoms prevented further root growth into deeper soil layers and induced proliferation of secondary roots, which confirmed the nematodes' pathogenicity. The severe infections in roots of corn plants suggest that parasitism of corn roots by the root-knot nematode must contribute to stunting, yellowing, and decline of corn, reducing yield by restricting access to water and nutrients that are needed for plant growth and development, and can result in the death of younger plants as previously reported (2). To our knowledge, this is the first report of M. incognita infecting corn in Spain. References: (1) K. R. Barker. Nematode extraction and bioassays. Page 19 in: An Advanced Treatise on Meloidogyne. Vol. II, Methodology. K. R. Barker et al., eds. North Carolina State University Graphics, Raleigh, 1985. (2) T. P. Heffes et al. Nematropica 22:139, 1992. (3) L. Robertson et al. Crop Prot. 25:440, 2006. (4) C. Zijlstra. Fund. Appl. Nematol. 20:505, 1997.

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First Report of Downy Mildew on Sweet Basil Caused By Peronospora belbahrii in India

Plant Disease ◽

10.1094/pdis-03-21-0621-pdn ◽

2021 ◽

Author(s):

Sujata Singh Yadav ◽

Priyanka Suryavanshi ◽

Indrajeet Nishad ◽

Soumya Sinha

Keyword(s):

Downy Mildew ◽

Disease Incidence ◽

Consensus Sequence ◽

Effective Control ◽

Broad Host Range ◽

Internal Transcribed Spacer Region ◽

Foliar Disease ◽

First Report ◽

Sweet Basil ◽

Initial Symptoms

Sweet basil (Ocimum basilicum L.; Family Lamiaceae) is an annual aromatic and medicinal plant grown in tropical and subtropical regions of the world. In India, it is cultivated as a commercial crop on ~8,000 ha. Aerial plant parts and essential oil of sweet basil are used in pharmaceutical, perfumery, food industries and in different formulations of traditional Ayurvedic and Unani medicines (Shahrajabian et al. 2020). The leaves have the highest concentrations of secondary metabolites such as terpenes and phenylpropanoids which provide the distinctive aroma (Viuda-Martos et al. 2011). During October 2020, severe foliar disease was observed in experimental fields of sweet basil at Council of Scientific and Industrial Research (CSIR)-Central Institute of Medicinal and Aromatic Plants (CIMAP) in Lucknow, India. Initial symptoms included large, interveinal chlorotic lesions on the adaxial surface of the leaves and black sporulation on the abaxial surface. Within a few days, the abaxial side of leaves turned necrotic, and leaf senescence and defoliation occurred on plants with severe symptoms. Disease incidence was 20 to 30% of plants. The pathogen was characterized morphologically using a light microscope. Sporangiophores were hyaline, dichotomously branched, 186.9 to 423.07 × 6.85 to 9.06 µm and, branched 3 to 5 times with each branch, terminating in two slightly curved branchlets, the longer one 7.05 to 25.31 µm and the shorter one 4.98 to 15.92 µm. Each branchlet had a single sporangium at the tip. Conidia were ellipsoidal to sub-globose, olive-brown in color, and typically measured 25.21 to 33.86 × 17.92 to 26.24 µm, each, without a pedicel. Based on these morphological characteristics, the foliar disease was identified as downy mildew was caused by Peronospora belbahrii (Thines et al. 2009). Eight symptomatic and two asymptomatic plant samples were collected from different locations in the field, and genomic DNA was extracted from the conidia of the eight naturally infected tissues of sweet basil samples as well as leaf tissues from two asymptomatic plants, using the CTAB method. The internal transcribed spacer region was amplified using ITS1 and ITS4 primers. Only eight infected samples amplified products of expected size (~ 700 bp) and two asymptomatic samples showed no amplification. Only five amplified PCR products were sequenced (White et al. 1990). All five sequences were identical and were a 98.1% match with five P. belbahrii isolates (MN450330.1, MN308051.1, MH620351.1, KJ960193, and MF693898). The consensus sequence was deposited into the NCBI database (GenBank Accession No. MW689257). Downy mildew caused by P. belbahrii previously has been reported on sweet basil from several countries (Wyenandt et al. 2015). To confirm the pathogenicity of these isolates on sweet basil (cv. CIM-Saumya), 25 - day-old sweet basil plants were sprayed with a suspension (1 × 105 sporangia/ml) of P. belbahrii. All plants were kept in a growth chamber with a 23/18°C diurnal cycle with 65 to 85% relative humidity for 24 h. Non-inoculated plants treated with sterile water served as a control treatment. After 8 days, typical symptoms of downy mildew appeared on all the inoculated plants while non-inoculated plants remained asymptomatic. Inoculated leaves with symptoms consistent of downy mildew were collected and the causal agent again identified as P. belbahrii on the basis of microscopic examination and ITS rDNA sequence data. To our knowledge, this is the first report of downy mildew caused by P. belbahrii on sweet basil in India. The pathogen has a broad host range and may pose a serious threat to the cultivation of this valuable crop in India. Thus, it is pertinent to develop effective control measures to avoid further spread and mitigate economic loss. References: Shahrajabian, M. H., et al. 2020. Int. J. Food Prop. 23:1961-1970. Wyenandt, C. A., et al. 2015. Phytopathology 105:885. Thines, M., et al. 2009. Mycol. Res. 113:532. White, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Viuda-Martos, M., et al. 2011. Food Control. 22:1715.

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